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We narrowed to 402 results for: PREP

Showing: 381 - 400 of 402 results
  1. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...AgeI. Mix: 6 μg pLKO.1 TRC-cloning vector (maxiprep or miniprep DNA) 5 μL 10x NEB buffer 1 1 μL AgeI add...use a miniprep kit to obtain DNA. 3. Conduct a restriction digest with EcoRI and NcoI: 1 μg miniprep DNA...Recommended Materials Material Vendor and catalog # DNA Miniprep Kit Qiagen #27104 EcoRI NEB #R0101S NcoI NEB #...based on your experiment Detailed protocols for preparing polybrene, protamine sulfate, and puromycin are... BL2 safety practices should be followed when preparing and handling lentiviral particles. Personal protective... 5g yeast extract 10g sodium chloride 15g agar Prepare LB agar solution by dissolving 40g of LB powder...store at 4°C. Hexadimethrine Bromide (Polybrene) Prepare a 1mg/mL solution of polybrene (Sigma-Aldrich catalog...
  2. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...BCD TFX) to 37 °C. See below for other Reagent Preparation instructions. Wipe down all pipettes and equipment...equipment with 10% bleach prior to use. Reagent Preparation 100 mM Valproic Acid Dissolve 2.88 g valproic...Sterilize by passage through a sterile 0.22 µm filter. Prepare 10 mL aliquots and store the solution at -20 °C... Filter the solution through a 0.22 µm filter. Prepare aliquots and store at -20 °C until use. BCD TFX... reagents Store at 4 °C until use. We suggest preparing fresh solutions after one month. BCD Feed 500 ... Glutagro Store at 4 °C until use. We suggest preparing fresh solutions after one month. 1000X protease...
  3. Pouring LB Agar Plates

    Type
    Protocol
    ...Use this protocol to prepare LB agar plates with antibiotic in your lab....samples are sterilizing in the autoclave, you should prepare your plate pouring station: Find an empty section...antibiotics, and a section for active pouring. Prepare your antibiotics. Prior to adding your antibiotic...1000x stock solution. For example: If you’ll be preparing plates with a final concentration of 100 ug/mL..., dissolve by vortexing, and filter sterilize. Prepare a water bath at 60 ℃ with sufficient water to submerge...Only the Resistant Strain Grows If everything was prepared properly, you should see growth of only the resistant...
  4. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...reagents to room temperature. Procedure Section 1: Prepare the Antigen Standard and coat the plate Dilute ... antigen to determine the ideal concentration. Prepare serial dilutions of the purified antigen as follows...overnight at 4 °C . Section 2: Block the plate Prepare the wash buffer (0.05% Tween-20 in PBS) as follows...wash, aspirate the wash buffer from the wells. Prepare the blocking buffer (1% BSA in PBS) as follows:...wash, aspirate the wash buffer from the wells. Prepare antibody dilution buffer (0.1% BSA in PBS) as follows...
  5. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...conditioned medium prepared above to make a new cell solution at a concentration of 5 cells/mL. Prepare approximately...culture dish, Corning 430167 (optional) Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-...a hemocytometer or other cell counter. Pro-Tip Prepare a relatively dilute cell solution (<10 6 cells/.... If you choose not to use conditioned medium, prepare this cell solution in fresh complete medium. This...
  6. Video Library

    Type
    Protocol
    ...Purification Purify adeno-associated virus from a preparation produced in mammalian cells AAV Purification ... Inoculating a Liquid Bacterial Culture How to prepare and grow bacteria in a liquid medium Inoculating...physical titer of your adeno-associated virus preparation. AAV Titration by qPCR Using SYBR Green Technology...
  7. General Transfection

    Type
    Protocol
    ... (PES) filter Syringes for filtering Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-...empirically determined for each new batch of 1 mg/mL PEI prepared. There may be variation between batches of PEI...empirically determined. Once a batch of PEI is prepared, transfect cells with a fluorescent plasmid using...
  8. Immunocytochemistry

    Type
    Protocol
    ...primary antibodies raised in a mouse. Reagent Preparation Permeabilization buffer: Dilute 20 µL of Triton...100 in 50 mL PBS. 300 nM DAPI working solution: Prepare a 300 µM DAPI stock solution by diluting 2.1 µL...DAPI solution to 100 µL PBS. Protect from light. Prepare a 300 nM DAPI working solution by diluting 5 µL...
  9. Lentivirus Production

    Type
    Protocol
    ...plasmid encoding your insert of interest Reagent Preparation DMEM Complete (also known as D10): 10% v/v FBS...Incubate the cells at 37 °C, 5% CO 2 for ~20 h. Prepare a mixture of the three transfection plasmids into...empirically determined. Once a batch of PEI is prepared, transfect cells with a fluorescent plasmid using...
  10. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...their resistance genes, as one does not need to prepare separate batches of antibiotic-containing agar....or other antibiotic resistance) Procedure Day 1 Prepare carbenicillin to a concentration of 1 mg/mL – 4...
  11. Centrifugation

    Type
    Protocol
    ...many different experimental procedures, such as minipreps and RNA extractions. Plus, centrifuges are useful...
  12. Guide to Using Pooled Libraries

    Type
    Guide
    ... simplest level, pooled libraries are single preparations of many different plasmids. Plasmids within ...either case, next-generation sequencing of the maxiprep DNA is recommended to verify that the library ...process: Amplify the library (electroporation and maxiprep). If delivering as virus, make virus; this creates...
  13. Using a Light Microscope Protocol

    Type
    Protocol
    ...image of that object! Equipment Light Microscope Prepared Slides (or other sample that can fit on a microscope...
  14. Kit Free RNA Extraction

    Type
    Protocol
    ...Protocol Before starting this protocol, make sure to prepare solution D (see reagent section above for the recipe...
  15. Adenovirus Guide

    Type
    Guide
    ...were discovered as a contaminant of adenovirus preparations, which is how they got their name, but they ...ensure no carryover from the HV in final viral preparations. Several methods to avoid HV contamination have...
  16. Science Guides

    Type
    Guide
    ...Pooled Libraries Pooled libraries are single preparations of many different plasmids. Plasmids within ...
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